GENETIC MECHANISMS IN TESTIS DIFFERENTIATION

睾丸分化的遗传机制

• 批准号: 2444949
• 负责人: VICKI N MEYERS-WALLEN
• 金额: $ 7.89万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-15 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2444949
• 关键词: animal genetic material tag; animal tissue; clone cells; developmental genetics; dogs; embryo /fetus cell /tissue; gene expression; genetic library; in situ hybridization; molecular cloning; nucleic acid sequence; polymerase chain reaction; sex differentiation; southern blotting; testis

DESCRIPTION (Adapted from applicant's description): The investigators plan to produce canine clones for genes that are known to be expressed in testis development and define the temporal and spatial expression patterns of these genes in normal canine embryonic gonads. The specific aims are: 1) To clone the entire coding sequence of canine SOX-9. The investigators are currently screening a canine genomic DNA library with a human SOX-9 that hybridizes to canine DNA. Homologous regions will be subcloned and characterized by sequence analysis. 2) To clone canine genomic DNA sequences of MIS (Amh), Wt-l, and SF-1. They will use standard library screening methods and polymerase chain reaction (PCR) to identify clones in a genomic DNA library. Regions of homology will be subcloned and their identity confirmed by sequence analysis. 3) To characterize the temporal and spatial expression patterns of Sry, Sox 9, MIS, Wt-1 and SF-1 in normal canine embryonic gonads, using reverse transcriptase PCR and in situ hybridization with canine probes. At the end of this study, they will be poised to examine the temporal and spatial expression patterns of these genes in affected embryos, which will be critical to understanding the genetic mechanisms responsible for abnormal testis development.

描述（改编自申请人的描述）：研究者计划 来克隆犬的睾丸中表达的基因 发展，并确定这些时空表达模式 正常犬胚胎性腺中的基因。 具体目标是：（1） 克隆犬SOX-9的全部编码序列。 调查人员正在 目前正在用人类SOX-9筛选犬基因组DNA文库， 与犬类DNA杂交 同源区域将被亚克隆， 其特征在于序列分析。 2)克隆犬基因组DNA MIS（Amh）、Wt-I和SF-1的序列。 他们将使用标准库 筛选方法和聚合酶链反应（PCR），以确定克隆， 基因组DNA文库。 同源性区域将被亚克隆，并且其 通过序列分析确认同一性。 3)为了描述时间的特征 Sry、Sox 9、MIS、Wt-1和SF-1在正常人中的表达及空间分布 犬胚胎性腺，使用逆转录酶PCR和原位 与犬探针杂交。 在本研究结束时，他们将 准备检查这些时间和空间的表达模式， 受影响的胚胎中的基因，这将是至关重要的了解， 导致睾丸发育异常的遗传机制。